水貂阿留申病毒荧光定量PCR检测方法的建立及初步应用  

Establishment and Application of Fluorescence Quantitative PCR Assay for Detection of Aleutian Mink Disease Virus

作　　者：张秀丽[1,2] 于慧娟 徐超[2] 张桉潮 宋兴超[2] 郝俊伟[1] 赵家平[2] 杜锐[1] 

机构地区：[1]吉林农业大学,吉林长春130118 [2]中国农业科学院特产研究所,吉林长春130122 [3]文登市泽库畜牧兽医工作站,山东文登264413 [4]吉林正大实业有限公司,吉林长春130033

出　　处：《中国畜牧兽医》2014年第9期1-5,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31272565)

摘　　要：根据水貂阿留申病毒(Aleutian mink disease virus,AMDV)基因序列(GenBank登录号:NC_001662.1)设计1对特异性引物,通过对荧光定量PCR反应条件的优化,建立了检测AMDV的SYBR GreenⅠ荧光定量PCR方法,该方法的灵敏度达10拷贝/μL,≥102拷贝/μL具有良好的特异性和重复性。同时利用该方法对8份疑似AMDV血清进行检测,结果6份阳性,阳性率为75%。本研究为水貂阿留申病的鉴别诊断及净群根除奠定了技术基础。One pair of specific primers was designed and synthesized according to the sequence of Aleutian mink disease vi- rus （AMDV） subbitted in the GenBank database, and then reaction requirements were optimized to develop a SYBR Green I fluorescence quantitative PCR assay. The results showed that the fluorescence quantitative PCR assay could detect 10 copies/μL of plasmid DNA. Its specificity and reproducibility were very good when 102 copies/μL. Meanwhile, 8 field sam- ples were detected with the methods and the positive rate was 75 %. This study would lay the foundation for differentiation di- agnosis and purification of the mink population.

关 键 词：水貂阿留申病毒 SYBR GreenⅠ 荧光定量PCR 标准曲线 

分 类 号：Q78[生物学—分子生物学]